title: PCR-RFLP identification of prohibited animal derived DNA in animal feed.
abstract: A method for confirming identification of prohibited species tissue in animal feed has been developed on the basis of PCR-RFLP analysis. In Japan, to prevent the spread of BSE through animal feed, the use of animal protein in feed has been regulated. Species-specific PCR detection of prohibited species materials in feed has been used as one of a series of laboratory tests to ensure the proper implementation of the feed regulations. However, since the result of this PCR method is determined only by amplicon length, it is sometimes necessary to confirm whether or not the positive result is due to the effect of a non-specific reaction. For this purpose, DNA sequencing is the best way to confirm the test result but it is not suitable for routine analysis because of the required time and cost. In this study, we developed an easy and rapid method to confirm the species identification (mammals, ruminants and cattle) by using 4 restriction enzymes: SmlI, MboI, BlnI and Hpy188III. This PCR-RFLP method, which ensures identification of prohibited animal species in feed, is useful for enhancing the reliability of feed inspection for BSE prevention. This method will be added to the Official Methods of Feed Analysis.
authors.affiliation: Food and Agricultural Materials Inspection Center, Chuo-ku, Saitama-shi, Saitama, Japan. naoki_shinoda@nm.famic.go.jp
authors.forename: Naoki
authors.initials: N
authors.lastname: Shinoda
authors.forename: Yoshiyasu
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: Masami
authors.initials: M
authors.lastname: Takagi
authors.forename: Fumio
authors.initials: F
authors.lastname: Kojima
authors.forename: Takashi
authors.initials: T
authors.lastname: Onodera
authors.forename: Katsuaki
authors.initials: K
authors.lastname: Sugiura
mesh_terms: D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D002417:Cattle; D004247:DNA / Q000032:analysis*; D016643:Encephalopathy, Bovine Spongiform / Q000517:prevention & control; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012418:Ruminants
chemical_list: D004247:DNA